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Application of Cryo-EM for Visualization of Mitoribosomes

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Mitochondrial Gene Expression

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2192))

Abstract

Mitochondrial ribosomes (mitoribosomes) are specialized machineries that carry out the synthesis of a limited number of proteins encoded in the mitochondrial genome, including components of the oxidative phosphorylation pathway. They have incorporated several structural features distinguishing them from bacterial and eukaryotic cytosolic counterparts. Our current understanding of the assembly and functioning of mitoribosomes is limited, and recent developments in cryo-EM provide promising directions for detailed investigation. Here we describe methods to purify mitoribosomes from human embryonic kidney cells for cryo-EM studies.

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Acknowledgements

We thank Juni Andréll and Shintaro Aibara for contributing to Figs. 2 and 6, respectively. This work was supported by the Swedish Foundation for Strategic Research (FFL15:0325), Ragnar Söderberg Foundation (M44/16), Swedish Research Council (NT_2015-04107), Cancerfonden (CAN 2017/1041), H2020-MSCA-ITN-2016 (REMIX). The cryo-EM data was collected at the Swedish national cryo-EM facility funded by the Knut and Alice Wallenberg, Family Erling Persson, and Kempe Foundations. We thank M. Carroni, J-M de la Rosa Trevin and S. Fleischmann for the smooth running of the data collection and processing.

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Authors and Affiliations

  1. Science for Life Laboratory, Department of Biochemistry and Biophysics, Stockholm University, Solna, Sweden

    Vivek Singh & Alexey Amunts

  2. Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden

    Vivek Singh & Alexey Amunts

Authors
  1. Vivek Singh
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  2. Alexey Amunts
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Correspondence to Alexey Amunts .

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Editors and Affiliations

  1. Medical Research Council Mitochondrial Biology Unit, University of Cambridge, Cambridge, UK

    Michal Minczuk

  2. Division of Molecular Metabolism, Department of Medical Biochemistry and Biophysics, Karolinska Institute, Solna, Sweden, Institutet Laboratory, Max Planck Institute Biology of Ageing - Karolinska, Karolinska Institutet, Stockholm, Sweden

    Joanna Rorbach

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Singh, V., Amunts, A. (2021). Application of Cryo-EM for Visualization of Mitoribosomes. In: Minczuk, M., Rorbach, J. (eds) Mitochondrial Gene Expression. Methods in Molecular Biology, vol 2192. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0834-0_15

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  • DOI: https://doi.org/10.1007/978-1-0716-0834-0_15

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0833-3

  • Online ISBN: 978-1-0716-0834-0

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